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In this study, the complete proteome of goat ejaculated semen including spermatozoa and seminal plasma was established, applying a tandem mass tag (TMT) labeling together with liquid chromatography-tandem mass spectrometry (LC-MS/MS). In seminal plasma, 2299 proteins were identified and 2098 proteins were quantified. The GO analysis demonstrated that 32% proteins were involved in metabolic activities. 46% proteins are located at intracellular region, intracellular organelle, and membrane-bounded organelle. Regarding molecular function, 40% proteins are engaged on protein binding, hydrolase activity, and ion binding. The KEGG analysis indicated a primary involvement of the identified proteins in protein processing in endoplasmic reticulum, lysosome, and proteome. In spermatozoa, 2491 proteins were identified and quantified. 39% proteins are involved in metabolic activities. 48% proteins are located at intracellular region, intracellular organelle, and membrane-bounded organelle. 38% proteins are engaged on protein binding, hydrolase activity, and ion binding. The KEGG analysis demonstrated their roles derived from the identified proteins in proteasome, glycolysis, pyruvate metabolism, and citrate cycle. Additionally, 1312 proteins were simultaneously presented in spermatozoa and seminal plasma. The involvement of 42% proteins in metabolic activities were observed. 47% proteins are located at intracellular region, intracellular organelle, and membrane-bounded organelle. The common proteins are mainly engaged on protein processing in endoplasmic reticulum, proteome, glycolysis, lysosome, and citrate cycle. Collectively, this study established the protein database of goat semen. More studies should be used to elucidate functionality of these identified proteins., (Copyright © 2022. Published by Elsevier Ltd.)

Phloem and xylem tissues are necessary for the allocation of nutrients and photoassimilates. However, how the long-distance transport of carbon (C) and nitrogen (N) is coordinated with the central metabolism is largely unknown. To better understand how the genetic and environmental factors influence C and N transport, we analysed the metabolite profiles of phloem exudates and xylem saps of five Arabidopsis thaliana accessions grown in low or non-limiting N supply. We observed that xylem saps were composed of 46 or 56% carbohydrates, 27 or 45% amino acids, and 5 or 13% organic acids in low or non-limiting N supply, respectively. In contrast, phloem exudates were composed of 76 or 86% carbohydrates, 7 or 18% amino acids, and 5 or 6% organic acids. Variation in N supply impacted amino acid, organic acid and sugar contents. When comparing low N and non-limiting N, the most striking differences were variations of glutamine, aspartate, and succinate abundance in the xylem saps and citrate and fumarate abundance in phloem exudates. In addition, we observed a substantial variation of metabolite content between genotypes, particularly under high N. The content of several organic acids, such as malate, citrate, fumarate, and succinate was affected by the genotype alone or by the interaction between genotype and N supply. This study confirmed that the response of the transport of nutrients in the phloem and the xylem to N availability is associated with the regulation of the central metabolism and could be an adaptive trait., (Copyright © 2022 Elsevier GmbH. All rights reserved.)

Red mature calyces of Hibiscus sabdariffa were collected from 16 different locations in Meghalaya, India. Samples were processed using shade drying (SD) and tray drying (TD). NMR spectroscopy was used to assess the metabolic composition of the calyces. In this study, 18 polar metabolites were assigned using 1D and 2D NMR spectra, and 10 of them were quantified. Proximate analysis showed that the TD method is more efficient at reducing moisture and maintaining the ash content of the Hibiscus biomass. NMR metabolomics indicates that the metabolite composition significantly differs between SD and TD samples and is more stable in TD plant processing. The differences in post-harvest drying has a greater impact on the metabolite composition of Hibiscus than the plant location.

Background: Exclusive breastfeeding is the optimal manner of early infant nutrition but women with gestational diabetes mellitus (GDM) often experience challenges with lactation in the early postpartum period. Increases in the colostral metabolites of lactose and citrate have been found to indicate increased milk production. Materials and Methods: A follow-up study of 133 postpartum women with and without GDM was conducted to examine differences in specific colostral metabolite levels using enzymatic methods to determine transition to lactogenesis II during the first week postpartum. We used linear mixed models for repeated measures over time to examine the effect of GDM on colostral metabolite levels at baseline and follow-up with fixed effects of GDM status, time, covariates, and interactions between time and GDM, between time and time, and between time, time and GDM into the model allowing quadratic trends over time. Results: Over time, lactose and citrate levels increased for all mothers ( p  < 0.001 and p  < 0.001, respectively), although mothers with GDM had consistently lower lactose and citrate levels compared with nondiabetic mothers ( p  = 0.004 and p  = 0.014, respectively). Age, prepregnancy body mass index, mode of birth, and parity did not independently influence colostral concentrations of lactose and citrate. Conclusions: Findings suggest that the rate of change overtime in lactose and citrate concentrations differ by GDM status. Further research examining the trajectory of colostral metabolite levels by GDM status is warranted.

Mass spectrometry imaging as a field has pushed its frontiers to three dimensions. Most three-dimensional mass spectrometry imaging (3D MSI) approaches require serial sectioning that results in a loss of biological information between analyzed slices and difficulty in reconstruction of 3D images. In this contribution, infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) was demonstrated to be applicable for 3D MSI that does not require sectioning because IR laser ablates material on a micrometer scale. A commercially available over-the-counter pharmaceutical was used as a model to demonstrate the feasibility of IR-MALDESI for 3D MSI. Depth resolution (i.e., z-resolution) as a function of laser energy levels and density of ablated material was investigated. The best achievable depth resolution from a pill was 2.3 μm at 0.3 mJ/pulse. 2D and 3D MSI were performed on the tablet to show the distribution of pill-specific molecules. A 3D MSI analysis on a region of interest of 15 × 15 voxels across 50 layers was performed. Our results demonstrate that IR-MALDESI is feasible with 3D MSI on a pill, and future work will be focused on analyses of biological tissues.

Existing spectrophotometric method to quantify hydroxycitric acid (HCA), although is specific and sensitive; finds limited use owing to poor stability of HCA-metavanadate complex. Present study describes improvisation of this method with respect to source of HCA standard and assay parameters. Assay system consisting of HCA and metavanadate reagent was modified to include 1 M NaOH to neutralize excess acidity. Resulting complex showed λ max at 485 nm, obeying Beer-Lambert's law within concentration range of 33-677 μg/ml, with linear calibration curve showing a good coefficient of determination (R 2  = 0.998). Moreover, HCA-metavanadate complex showed enhanced stability retaining up to 70% absorbance even after 60 min of its formation. Similar consistency in scaled-down assay system renders the method suitable for high-throughput screening of HCA-producing microbes. Of the tested metabolites and media components, only tartrate interfered with the spectrophotometric estimation of HCA; a correction factor to eliminate which was also established. Accordingly measured HCA level in the culture supernatant of a bacterial isolate IT6 was comparable to that determined using the standardized HPLC method. The proposed procedure therefore is a convenient, sensitive, accurate and high-throughput method suitable for primary screening of HCA producing microbes; the only ecofriendly alternative source of optically pure HCA., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Siderophores, iron-scavenging small molecules, are fundamental to bacterial nutrient metal acquisition and enable pathogens to overcome challenges imposed by nutritional immunity. Multimodal imaging mass spectrometry allows visualization of host-pathogen iron competition, by mapping siderophores within infected tissue. We have observed heterogeneous distributions of Staphylococcus aureus siderophores across infectious foci, challenging the paradigm that the vertebrate host is a uniformly iron-depleted environment to invading microbes., Competing Interests: The authors declare no competing interest., (Copyright © 2019 the Author(s). Published by PNAS.)

Phosphorus-based compounds are used as plasticizers in the manufacture of many products found in the indoor environment. Here we quantitatively investigated dermal exposure to phosphorus-based compounds contained in 45 nail polishes purchased in Japan. The alternative plasticizer triphenyl phosphate (TPhP) was detected in some samples of the nail polishes made in the USA (concentration, 1.1-1.8 wt%). The potential dermal exposure rates for TPhP, estimated using ConsExpo (version 5.0; Dutch National Institute for Public Health and the Environment), were in the range 200 (5%ile)-1700 (50%ile)-5000 (95%ile) ng kg-bw -1 day -1 , which is more than 1400 times the reported values for exposure via dust ingestion and inhalation. Thus, dermal exposure via nail polish may be a major route of exposure to TPhP. The margin of exposure range for TPhP was 3.6 × 10 5 -4.1 × 10 4 -1.4 × 10 4 . For comparison, the potential dermal exposure rate range for the conventional plasticizer dibutyl phthalate and the alternative plasticizer acetyl tributyl citrate was 360-3500-14,000 and 430-4100-17,000 ng kg-bw -1 day -1 , respectively, and the margin of exposure range was 4.1 × 10 3 -4.2 × 10 2 -1.1 × 10 2 and 2.3 × 10 5 -2.4 × 10 4 -5.9 × 10 3 , respectively., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Gastrodia elata ( G. elata ) tuber is a valuable herbal medicine used to treat many diseases. The procedure of establishing a reasonable and feasible quality assessment method for G. elata tuber is important to ensure its clinical safety and efficacy. In this research, an effective and comprehensive evaluation method for assessing the quality of G. elata has been developed, based on the analysis of high performance liquid chromatography (HPLC) fingerprint, combined with the quantitative analysis of multi-components by single marker (QAMS) method. The contents of the seven components, including gastrodin, p -hydroxybenzyl alcohol, p -hydroxy benzaldehyde, parishin A, parishin B, parishin C, and parishin E were determined, simultaneously, using gastrodin as the reference standard. The results demonstrated that there was no significant difference between the QAMS method and the traditional external standard method (ESM) ( p > 0.05, RSD < 4.79%), suggesting that QAMS was a reliable and convenient method for the content determination of multiple components, especially when there is a shortage of reference substances. In conclusion, this strategy could be beneficial for simplifying the processes in the quality control of G. elata tuber and giving references to promote the quality standards of herbal medicines.

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by systemic, symmetrical, and erosive synovitis. RA is one of the most common disabling diseases in the clinic. The main clinical intervention strategies are early diagnosis and early treatment. This study aims to predict the diagnostic value of 68 Ga-citrate and 18 F-FDG PET/CT in RA by comparing and analyzing the value of 68 Ga-citrate and 18 F-FDG PET/CT for diagnosing type II collagen-induced arthritis (CIA) in rats. Some CIA models were established. Normal rats were selected as the control group, and 23 days and 40 days were selected as the early and late time points of arthritis, respectively. The semiquantitative analysis of CIA rats was carried out with 68 Ga-citrate PET/CT and 18 F-FDG PET/CT, and the ratio of the maximum standardized uptake (SUV max ) values in the regions of interest (ROIs) of the hind foot ankle joint and thigh muscle was calculated and statistically analyzed. The distribution of CIA rats in vivo at the 68 Ga-citrate 90 min time point was studied, and the ankle tissues were evaluated with hematoxylin and eosin (HE) staining. 68 Ga-citrate PET/CT is obviously superior to 18 F-FDG PET/CT for CIA imaging, and the statistical results show that the difference between the two examination methods is statistically significant ( P < 0.001). The uptake of these two radiopharmaceuticals showed the same trend in arthritis rats with different scores. The distribution of 68 Ga-citrate at 90 min is consistent with the trend shown by 68 Ga-citrate PET/CT. 68 Ga-citrate PET/CT can reflect the inflammatory activity of affected joints in CIA rats earlier and more sensitively than 18 F-FDG PET/CT, and this imaging advantage continues until the later stage of inflammation. Therefore, 68 Ga-citrate PET/CT is worthy of further promotion and application in the clinical diagnosis of RA.

Introduction: Lactogenesis II (LaII) failure can be prevented in at-risk mothers with simple proactive interventions. In a randomised trial, we investigated the efficacy of early and regular breast milk expression in establishing LaII, using an electric double-breast pump., Methods: Mothers with uncomplicated singleton deliveries were randomised to intervention (n = 31) or control (n = 29) groups. The former commenced breast milk expression with an electric pump within one hour of delivery and maintained regular expression with direct breastfeeding. Control mothers directly breastfed without regular pump expression. Expressed milk volumes were analysed for citrate, lactose, sodium and protein., Results: Median time of LaII was Day 3 (interquartile range [IQR] 1 day) with intervention and on Day 4 (IQR 1 day) among controls (p = 0.03). Biochemical steady-state concentrations were achieved around early Day 4 (sodium, total protein) and Days 4-5 (citrate, lactose). Sodium, protein and lactose levels were similar in both groups over seven days, at 5.80 mM, 0.68 mM and -13.38 mM, respectively. Mean daily milk volume with intervention was 73.9 mL on Day 3 and 225.2 mL on Day 7, greater than controls (25.4 mL on Day 3 and 69.2 mL on Day 7; p < 0.2). Mean infant weights were similar on Day 8 at 3,477 g with intervention and 3,479 g among controls., Conclusion: LaII is established by postnatal Day 3 with early initiation of regular breast milk expression, a useful intervention for mothers at risk of early-onset breastfeeding failure., (Copyright: © Singapore Medical Association.)

Garcinia L. (Clusiaceae) fruits are a rich source of (-)-hydroxycitric acid, and this has gained considerable attention as an anti-obesity agent and a popular weight loss food supplement. In this study, we assessed adulteration of morphologically similar samples of Garcinia using DNA barcoding, and used NMR to quantify the content of (-)-hydroxycitric acid and (-)-hydroxycitric acid lactone in raw herbal drugs and Garcinia food supplements. DNA barcoding revealed that mostly G. gummi-gutta (previously known as G. cambogia) and G. indica were traded in Indian herbal markets, and there was no adulteration. The content of (-)-hydroxycitric acid and (-)-hydroxycitric acid lactone in the two species varied from 1.7% to 16.3%, and 3.5% to 20.7% respectively. Analysis of ten Garcinia food supplements revealed a large variation in the content of (-)-hydroxycitric acid, from 29 mg (4.6%) to 289 mg (50.6%) content per capsule or tablet. Only one product contained quantifiable amounts of (-)-hydroxycitric acid lactone. Furthermore the study demonstrates that DNA barcoding and NMR could be effectively used as a regulatory tool to authenticate Garcinia fruit rinds and food supplements.

Plastic materials are widely used in food packaging applications; however, there is increased concern because of the possible release of undesirable components into foodstuffs. Migration of plastic constituents not only has the potential to affect product quality but also constitutes a risk to consumer health. In order to check the safety of food contact materials, analytical methodologies to identify potential migrants are required. In the first part of this work, a GC/MS screening method was developed for the identification of components from plastic packaging materials including intentionally and "non-intentionally added substances" (NIAS) as potential migrants. In the second part of this study, the presence of seven compounds (bis (2-ethylhexyl) phthalate (DEHP), diethyl phthalate (DEP), diisobutyl phthalate (DIBP), dibutyl phthalate (DBP), butylated hydroxytoluene (BHT), acetyl tributyl citrate (ATBC), benzophenone (BP)) previously identified in packaging materials were investigated in food products (corn and potatoes snacks, cookies, and cakes). For this purpose, a suitable extraction method was developed and quantification was performed using GC-MS. The developed method was validated in terms of linearity, recovery, repeatability, and limits of detection and quantification. The spiked recoveries varied between 82.7 and 116.1%, and relative standard deviation (RSD) was in the range of 2.22-15.9%. The plasticizer ATBC was the most detected compound (94% samples), followed by DEP (65%), DEHP (47%), BP (44%), DBP (35%), DIBP (21%), and BHT (12%). Regarding phthalates, DEP and DEHP were the most frequently detected compounds in concentrations up to 1.44 μg g -1 . In some samples, only DBP exceeded the European SML of 0.3 mg kg -1 established in Regulation 10/2011. Graphical abstract Chemical migration from plastic packaging into food.

Objectives: The purpose was to evaluate a combined effect of active ingredients contained in new toothpaste in reducing dentine hypersensitivity., Methods: The tested toothpaste contained active ingredients potassium nitrate (5.00%) and zinc citrate (0.50%). Pain to water stimulus and blowing air was reported on hypersensitive teeth and measured using the VAS (Visual Analogue Scale). The degree of pain to water stimulus was detected after each subject drank a few sips of water which was boiled to the room temperature. Data were collected on each subject's dental history., Results: A significant reduction in dentine hypersensitivity (26-29% after 2 weeks versus 40% after 4 weeks) was detected in the tested group. The controls also showed significant reduction to both stimuli, but at lower rate. A statistically significant difference to blowing air and water stimuli showed the highest impact for the tested group after 4 weeks (P = 0.033, P = 0.006)., Conclusion: This study supports the efficacy of the tested toothpaste in reducing the phenomenon of dentine hypersensitivity after 4 weeks of usage., (© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objective: To determine whether intermittent hypoxia (IH) persisting after 36 weeks postmenstrual age (PMA) can be attenuated using caffeine doses sufficient to maintain caffeine concentrations >20 μg ml -1 ., Study Design: Twenty-seven infants born <32 weeks were started on caffeine citrate at 10 mg kg -1  day -1 when clinical caffeine was discontinued. At 36 weeks PMA, the dose was increased to 14 or 20 mg kg -1  day -1 divided twice a day (BID) to compensate for progressively increasing caffeine metabolism. Caffeine concentrations were measured weekly. The extent of IH derived from continuous pulse oximetry was compared to data from 53 control infants., Result: The mean (s.d.) gestational age of enrolled infants was 27.9±2 weeks. Median caffeine levels were >20 μg ml -1 on study caffeine doses. IH was significantly attenuated through 38 weeks PMA compared with the control group., Conclusion: Caffeine doses of 14 to 20 mg kg -1  day -1 were sufficient to maintain caffeine concentrations >20 μg ml -1 and reduce IH in preterm infants at 36 to 38 weeks PMA.

Chronic constipation and gastrointestinal motility disorders constitute a large part of a gastroenterology practice and have a significant impact on a patient's quality of life and lifestyle. In most cases, medications are prescribed to alleviate symptoms without there being an objective measurement of response. Commonly used investigations of gastrointestinal transit times are currently limited to radiopaque markers or electronic capsules. Repeated use of these techniques is limited because of the radiation exposure and the significant cost of the devices. We present the proof of concept for a new device to measure gastrointestinal transit time using commonly available and inexpensive materials with only a small amount of radiotracer. Methods: We assembled gelatin capsules containing a 67 Ga-citrate-radiolabeled grain of rice embedded in paraffin for use as a point-source transit device. It was tested for stability in vitro and subsequently was given orally to 4 healthy volunteers and 10 patients with constipation or diarrhea. Imaging was performed at regular intervals until the device was excreted. Results: The device remained intact and visible as a point source in all subjects until excretion. When used along with a diary of bowel movement times and dates, the device could determine the total transit time. The device could be visualized either alone or in combination with a barium small-bowel follow-through study or a gastric emptying study. Conclusion: The use of a point-source transit device for the determination of gastrointestinal transit time is a feasible alternative to other methods. The device is inexpensive and easy to assemble, requires only a small amount of radiotracer, and remains inert throughout the gastrointestinal tract, allowing for accurate determination of gastrointestinal transit time. Further investigation of the device is required to establish optimum imaging parameters and reference values. Measurements of gastrointestinal transit time may be useful in managing patients with dysmotility and in selecting the appropriate pharmaceutical treatment., (© 2017 by the Society of Nuclear Medicine and Molecular Imaging.)

The growing production and commercial application of engineered nanoparticles (ENPs), such as Ag, CeO 2 , and TiO 2 nanoparticles, induce a risk to the environment as ENPs are released during their use. The comprehensive assessment of the environmental risk that the ENPs pose involves understanding their fate and behavior in wastewater treatment systems. Therefore, in this study, we investigate the effect of plants and different substrates on the retention and distribution of citrate-coated silver nanoparticles (Ag-NPs) in batch experimental setups simulating constructed wetlands (CWs). Sand, zeolite, and biofilm-coated gravel induce efficient removal (85, 55, and 67 %, respectively) of Ag from the water phase indicating that citrate-coated Ag-NPs are efficiently retained in CWs. Plants are a minor factor in retaining Ag as a large fraction of the recovered Ag remains in the water phase (0.42-0.58). Most Ag associated with the plant tissues is attached to or taken up by the roots, and only negligible amounts (maximum 3 %) of Ag are translocated to the leaves under the applied experimental conditions.

The genus Garcinia shows a considerable variation in its morphological characters such as leaf, flower and fruit with taxonomic ambiguity. It is a potential under-exploited multipurpose crop that gained considerable attention for the presence of (-) hydroxycitric acid, an anti-obesity compound, in its fruit rind and leaves. Here, we evaluated the genetic relationship through molecular markers among the selected 9 species commonly available in the Western Ghats and the Northeastern Himalayan foot hills of India. The nucleotide sequence data obtained from two prominent monomorphic bands generated in ISSR profiling of the species was utilized for the study. The selected bands were found to be of ITS region (700 bp) and partial region of KNOX-1 gene (600 bp). The evolutionary cluster was formed using MEGA5 software. The study indicated 2 major clusters, influenced by floral morphology of the species and availability of (-) hydroxycitric acid in their fruit rinds. In the subclusters, one species from the Western Ghats were paired with another from Northeastern Himalayas with relatively similar morphological traits.

Edwardsiella tarda has long been known as a pathogen that causes severe economic losses in aquaculture industry. Insights gained on E. tarda pathogenesis may prove useful in the development of new methods for the treatment of infections as well as preventive measures against future outbreaks. In this report, we have established the correlation between the presence of virulence genes, related with three aspects typically involved in bacterial pathogenesis (chondroitinase activity, quorum sensing and siderophore-mediated ferric uptake systems), in the genome of E. tarda strains isolated from turbot in Europe and their phenotypic traits. A total of 8 genes were tested by PCR for their presence in 73 E. tarda isolates. High homogeneity was observed in the presence/absence pattern of all the strains. Positive results in the amplification of virulence-related genes were correlated with the detection of chondroitinase activity in agar plates, in vivo AHL production during fish infection and determination of type of siderophore produced by E. tarda. To the best of our knowledge, this is the first study carried out with European strains on potential virulence factors. Furthermore, we demonstrated for the first time that E. tarda produces the siderophore vibrioferrin., (© 2015 John Wiley & Sons Ltd.)

This study evaluated the influence of parameters such as temperature and type of low-density polyethylene (LDPE) film on the log Kp/f values of seven model migrants in food simulants. Two different types of LDPE films contaminated by extrusion and immersion were placed in contact with three food simulants including 20% ethanol, 50% ethanol and olive oil under several time-temperature conditions. Results suggest that most log Kp/f values are little affected by these parameters in this study. In addition, the relation between log Kp/f and log Po/w was established for each food simulant and regression lines, as well as correlation coefficients, were calculated. Correlations were compared with data from real foodstuffs. Data presented in this study could be valuable in assigning certain foods to particular food simulants as well as predicting the mass transfer of potential migrants into different types of food or food simulants, avoiding tedious and expensive laboratory analysis. The results could be especially useful for regulatory agencies as well as for the food industry.

Background: Iron fortification of rice is a promising strategy for improving iron nutrition. However, it is technically challenging because rice is consumed as intact grains, and ferric pyrophosphate (FePP), which is usually used for rice fortification, has low bioavailability., Objective: We investigated whether the addition of a citric acid/trisodium citrate (CA/TSC) mixture before extrusion increases iron absorption in humans from FePP-fortified extruded rice grains., Design: We conducted an iron absorption study in iron-sufficient young women (n = 20), in which each participant consumed 4 different meals (4 mg Fe/meal): 1) extruded FePP-fortified rice (No CA/TSC); 2) extruded FePP-fortified rice with CA/TSC added before extrusion (CA/TSC extruded); 3) extruded FePP-fortified rice with CA/TSC solution added after cooking and before consumption (CA/TSC solution); and 4) nonextruded rice fortified with a FeSO4 solution added after cooking and before consumption (reference). Iron absorption was calculated from erythrocyte incorporation of stable iron isotopes 14 d after administration. In in vitro experiments, we assessed the soluble and dialyzable iron from rice meals in which CA/TSC was added at different preparation stages and from meals with different iron:CA:TSC ratios., Results: Fractional iron absorption was significantly higher from CA/TSC-extruded meals (3.2%) than from No CA/TSC (1.7%) and CA/TSC solution (1.7%; all P < 0.05) and was not different from the FeSO4 reference meal (3.4%). In vitro solubility and dialyzability were higher in CA/TSC-extruded rice than in rice with No CA/TSC and CA/TSC solution, and solubility increased with higher amounts of added CA and TSC in extruded rice., Conclusions: Iron bioavailability nearly doubled when CA/TSC was extruded with FePP into fortified rice, resulting in iron bioavailability comparable to that of FeSO4 We attribute this effect to an in situ generation of soluble FePP citrate moieties during extrusion and/or cooking because of the close physical proximity of FePP and CA/TSC in the extruded rice matrix. This trial was registered at clinicaltrials.gov as NCT02176759., (© 2016 American Society for Nutrition.)

The ability to detect radiation in microfluidic devices is important for the on-chip analysis of radiopharmaceuticals, but previously reported systems have largely suffered from various limitations including cost, complexity of fabrication, and insufficient sensitivity and/or speed. Here, we present the use of sensitive, low cost, small-sized, commercially available silicon photomultipliers (SiPMs) for the detection of radioactivity inside microfluidic channels fabricated from a range of conventional microfluidic chip substrates. We demonstrate the effects of chip material and thickness on the detection of the positron-emitting isotope, [(18)F]fluoride, and find that, while the SiPMs are light sensors, they are able to detect radiation even through opaque chip materials via direct positron and gamma (γ) ray interaction. Finally, we employed the SiPM platform for analysis of the PET (positron emission tomography) radiotracers 2-[(18)F]fluoro-2-deoxy-d-glucose ([(18)F]FDG) and [(68)Ga]gallium-citrate, and highlight the ability to detect the γ ray emitting SPECT (single photon emission computed tomography) radiotracer, [(99m)Tc]pertechnetate.

Food emulsifiers are widely used to stabilise water-fat emulsions such as mayonnaise and dressings. They are prepared by oligomerisation of a poly-alcohol (as e.g. glycerol or citric acid) followed by a reaction with fatty acids. In order to gain insight in the chemical composition of different emulsifiers, a range of chromatographic methods including gas chromatography, size exclusion chromatography, normal phase- and reversed phase liquid chromatography either or not in combination with mass spectrometry was deployed. The different methods turned out to be highly complementary. By combining the information from different methods the polar head group and the fatty acid part of the emulsifier can be characterised in detail. Mass spectrometry is indispensable for establishing the number of polar molecules in the head group as well as for establishing the correct combinations of fatty acids in one molecule. Ten commercial emulsifiers were described at the level of number and type of polar groups and fatty acids present., (Copyright © 2016 Elsevier B.V. All rights reserved.)

This work reports two new diketoprrrolopyrrole-based fluorescent chemosensors (DPP-Py1 and DPP-Py2) using symmetrical diamides as recognition groups for selective and fast detection of citrate in the near-infrared region. To our delight, DPP-Py1 is a ratiometric sensor, whereas DPP-Py2 is a turn-on fluorescent sensor. It is worth noting that DPP-Py1 has higher accuracy and sensitivity with a relatively lower detection limit (1.8 × 10(-7) M) and better stability in different pH buffers than DPP-Py2. Scanning electron microscopy, dynamic light scattering analyses, (1)H NMR titration, and 2D-NOESY NMR suggested that the fluorescence increment of the probes DPP-Py1 and DPP-Py2 for citrate could probably originate from aggregation-induced emission (AIE) on the basis of the complexation of the pyridinium-based symmetrical diamides, DPPs, with carboxyl anions of citrate. Our work may provide a simpler and faster means for qualitative and quantitative analysis of citrate through an AIE mechanism.

A simple, fast, precise, economic, selective and accurate HPLC method for simultaneous estimation of sorbicacid, sodium picosulphate and methyl parabensodium in laxative drops has been developed and subsequently validated. Chromatographic separation was achieved using gradient elution with mix phosphate buffer pH 7.0 and acetonitrile. The column used was purospherstar C18, 5 μm, 25 cm × 4.6mm kept at 25°C with 1 ml/min flow rate using detection (PDA) at 263 nm. The retention times of sorbicacid, sodium picosulphate and methyl paraben sodium were found to be 4.6, 7.4 and 11.4 minutes respectively. The proposed method was found to be linear over a concentration range of 8-12 μg/ml for sorbic acid, 60-90 μg/ml for sodium picosulphate and 16-24 μg/ml formethyl paraben sodium respectively. The recovery was found to be 99.13-101.68% for sorbic acid, 99.81-100.21% for sodium picosulphate and 99.84-100.09% for methyl paraben sodium respectively. The limit of detection (LOD) for sorbicacid, sodium picosulphate and methyl parabensodium were found to be 0.032 μg/ml, 0.337 μg/ml and 0.131 μg/ml respectively and limit of quantitation (LOQ) for sorbicacid, sodium picosulphate and methyl parabensodium were found to be 0.097 μg/ml, 1.023 μg/ml and 0.399 μg/ml respectively. The method was validated with respect to specificity, precision, accuracy, linearity and robustness according to guidelines of ICH.

Electronic tongue technology based on arrays of cross-sensitive chemical sensors and chemometric data processing has attracted a lot of researchers' attention through the last years. Several so far reported applications dealing with pharmaceutical related tasks employed different e-tongue systems to address different objectives. In this situation, it is hard to judge on the benefits and drawbacks of particular e-tongue implementations for R&D in pharmaceutics. The objective of this study was to compare the performance of six different e-tongues applied to the same set of pharmaceutical samples. For this purpose, two commercially available systems (from Insent and AlphaMOS) and four laboratory prototype systems (two potentiometric systems from Warsaw operating in flow and static modes, one potentiometric system from St. Petersburg, one voltammetric system from Barcelona) were employed. The sample set addressed in the study comprised nine different formulations based on caffeine citrate, lactose monohydrate, maltodextrine, saccharin sodium and citric acid in various combinations. To provide for the fair and unbiased comparison, samples were evaluated under blind conditions and data processing from all the systems was performed in a uniform way. Different mathematical methods were applied to judge on similarity of the e-tongues response from the samples. These were principal component analysis (PCA), RV' matrix correlation coefficients and Tuckeŕs congruency coefficients., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Manufactured metallic silver nanoparticles (AgNP) are intensively utilized in consumer products and this will inevitably lead to their release to soils. To assess the environmental risks of AgNP in soils, quantification of both their concentration and size in soil solution is essential. We developed a methodology consisting of asymmetric flow field-flow fractionation (AF4) in combination with on-line detection by UV-vis spectroscopy and off-line HR-ICP-MS measurements to quantify the concentration and size of AgNP, coated with either citrate or polyvinylpyrrolidone (PVP), in water extracts of three different soils. The type of mobile phase was a critical factor in the fractionation of AgNP by AF4. In synthetic systems, fractionation of a series of virgin citrate- and PVP-coated AgNP (10-90 nm) with reasonably high recoveries could only be achieved with ultrahigh purity water as a mobile phase. For the soil water extracts, 0.01% (w:v) sodium dodecyl sulfate (SDS) at pH 8 was the key to a successful fractionation of the AgNP. With SDS, the primary size of AgNP in all soil water extracts could be determined by AF4, except for PVP-coated AgNP when clay colloids were present. The PVP-coated AgNP interacted with colloidal clay minerals, leading to an overestimation of their primary size. Similar interactions between PVP-coated AgNP and clay colloids can take place in the environment and facilitate their transport in soils, aquifers, and surface waters. In conclusion, AF4 in combination with UV-vis spectroscopy and HR-ICP-MS measurements is a powerful tool to characterize AgNP in soil solution if the appropriate mobile phase is used., (Copyright © 2015 Elsevier B.V. All rights reserved.)

An ultra performance liquid chromatography (UHPLC) coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF-MS/MS) was used in the structural determination of natural compounds in Gastrodia elata. A total of 64 compounds were identified or tentatively characterized. The strategy used for characterization was comparing their retention time and fragmentation behaviors with those of the reference standards, or investigating their accurate mass measurements and characteristic fragmentation patterns followed by low-energy collision dissociation tandem mass spectrometry (CID-MS/MS). Phenolic conjugates mainly underwent consecutive losses of gastrodin residues and combined losses of H2O and CO2 from their citric acid units under negative MS/MS conditions. According to these rules, we have successfully characterized fifteen potential novel compounds. To confirm the reliability of this strategy, two targeted unknown trace parishins were obtained from G. elata by LC/MS-guided isolation. Based on the analysis of data from NMR spectroscopy and other techniques, the two unknown parishins were identified as 2-[4-O-(β-d-glucopyranosyl)benzyl]-3-methyl-citrate (parishin J) and 1,2-di-[4-O-(β-d-glucopyranosyl)benzyl]-3-methyl-citrate (parishin K), respectively. The fully established structures were consistent with the MS-oriented structural elucidation. This study expanded our knowledge on parishins in Gastrodia species, and the proposed strategy was proven efficient and reliable in the discovery of new minor compounds from herbal extracts., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The binding constant determination of uranyl with small-molecule ligands such as citric acid could provide fundamental knowledge for a better understanding of the study of uranyl complexation, which is of considerable importance for multiple purposes. In this work, the binding constant of uranyl-citrate complex was determined by ACE. Besides the common single-injection method, a multi-injection method to measure the electrophoretic mobility was also applied. The BGEs used contained HClO4 and NaClO4 , with a pH of 1.98 ± 0.02 and ionic strength of 0.050 mol/L, then citric acid was added to reach different concentrations. The electrophoretic mobilities of the uranyl-citrate complex measured by both of the two methods were consistent, and then the binding constant was calculated by nonlinear fitting assuming that the reaction had a 1:1 stoichiometry and the complex was [(UO2 )(Cit)](-) . The binding constant obtained by the multi-injection method was log K = 9.68 ± 0.07, and that obtained by the single-injection method was log K = 9.73 ± 0.02. The results provided additional knowledge of the uranyl-citrate system, and they demonstrated that compared with other methods, ACE using the multi-injection method could be an efficient, fast, and simple way to determine electrophoretic mobilities and to calculate binding constants., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Citric acid, a widespread soil rhizosphere plant/microbe carboxylic acid exudate can easily form chelates with heavy metals, increasing their availability in the environment. When Cu(II) from algal control in water bodies or reservoirs and fungicides, such as the Bordeaux mixture, and citrate interact, solubilization through chelation is a possible outcome. Manganese (hydr)oxides represent a significant portion of the subsurface environment and can affect the fate and transport of chemical species through adsorption and oxidation. This study explores the possible interaction between MnO2 and Cu-Citrate under ambient oxic conditions. The calculated Mn(II) dissolution rates during the initial 1h of reaction followed the series Cu(II)>Cu-Citrate 1:0.5>Cu-Citrate 1:1(oxic)>Citrate>Cu-Citrate 1:1(Anoxic), reinforcing the central role of (complexed or un-complexed) Cu(II) during the initial surface-coordination instead of following the s-shaped auto-catalytic curves of Mn(II) dissolution in citrate solution. The use of capillary electrophoresis allowed the detection of an intermediate Cu(II)Acetonedicarboxylate complex and the oxidation products acetonedicarboxylate, acetoacetate, acetone and acetic acid. The mass balance analysis of Cu-Citrate 1:1 suggests the partial adsorption of Cu-Citrate(ads) and catalytic degradation of acetonedicarboxylate through a MnO2-Cu surface sorbed complex. Lastly, XPS analysis confirmed the MnO2 surface Cu(II) reduction along with an outer-hydration layer at the MnO2 interface, where electron transfer and aquo ligand exchange may lead to the oxidation of Cu-Citrate., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Spectral degradations as a result of temporal field variations are observed in MRSI of the human prostate. Moving organs generate substantial temporal and spatial field fluctuations as a result of susceptibility mismatch with the surrounding tissue (i.e. periodic breathing, cardiac motion or random bowel motion). Nine patients with prostate cancer were scanned with an endorectal coil (ERC) on a 7-T MR scanner. Temporal B0 field variations were observed with fast dynamic B0 mapping in these patients. Simulations of dynamic B0 corrections were performed using zero- to second-order shim terms. In addition, the temporal B0 variations were applied to simulated MR spectra causing, on average, 15% underestimation of the choline/citrate ratio. Linewidth distortions and frequency shifts (up to 30 and 8 Hz, respectively) were observed. To demonstrate the concept of observing local field fluctuations in real time during MRSI data acquisition, a field probe (FP) tuned and matched for the (19)  F frequency was incorporated into the housing of the ERC. The data acquired with the FP were compared with the B0 field map data and used to correct the MRSI datasets retrospectively. The dynamic B0 mapping data showed variations of up to 30 Hz (0.1 ppm) over 72 s at 7 T. The simulated zero-order corrections, calculated as the root mean square, reduced the standard deviation (SD) of the dynamic variations by an average of 41%. When using second-order corrections, the reduction in the SD was, on average, 56%. The FP data showed the same variation range as the dynamic B0 data and the variation patterns corresponded. After retrospective correction, the MRSI data showed artifact reduction and improved spectral resolution. B0 variations can degrade the MRSI substantially. The simple incorporation of an FP into an ERC can improve prostate cancer MRSI without prior knowledge of the origin of the dynamic field distortions., (Copyright © 2014 John Wiley & Sons, Ltd.)

The chemical synthesis of staphyloferrin A, a siderophore used by Staphylococcus bacteria for ferric iron retrieval, has been achieved with 79% yield via solid phase peptide synthesis (SPPS). Biological activity of synthetic staphyloferrin A has been confirmed by demonstrating its capture and uptake by live S. aureus.

Today most foods are available in a packed form. During storage, the migration of chemical substances from food packaging materials into food may occur and may therefore be a potential source of consumer exposure. To protect the consumer, standard migration tests are laid down in Regulation (EU) No. 10/2011. When using those migration tests and applying additional conservative conventions, estimated exposure is linked with large uncertainties including a certain margin of safety. Thus the research project FACET was initiated within the 7th Framework Programme of the European Commission with the aim of developing a probabilistic migration modelling framework which allows one (1) to calculate migration into foods under real conditions of use; and (2) to deliver realistic concentration estimates for consumer exposure modelling for complex packaging materials (including multi-material multilayer structures). The aim was to carry out within the framework of the FACET project a comprehensive systematic study on the solubility behaviour of foodstuffs for potentially migrating organic chemicals. Therefore a rapid and convenient method was established to obtain partition coefficients between polymer and food, KP/F. With this method approximately 700 time-dependent kinetic experiments from spiked polyethylene films were performed using model migrants, foods and ethanol-water mixtures. The partition coefficients of migrants between polymer and food (KP/F) were compared with those obtained using ethanol-water mixtures (KP/F's) to investigate whether an allocation of food groups with common migration behaviour to certain ethanol-water mixtures could be made. These studies have confirmed that the solubility of a migrant is mainly dependent on the fat content in the food and on the ethanol concentration of ethanol-water mixtures. Therefore dissolution properties of generic food groups for migrants can be assigned to those of ethanol-water mixtures. All foodstuffs (including dry foods) when allocated to FACET model food group codes can be classified into a reduced number of food categories each represented by a corresponding ethanol-water equivalency.

A microfluidic bioreactor with an easy to fabricate nano-plasmonic surface is demonstrated for studies of biofilms and their precursor materials via Surface Enhanced Raman Spectroscopy (SERS). The system uses a novel design to induce sheath flow confinement of a sodium citrate biofilm precursor stream against the SERS imaging surface to measure spatial variations in the concentration profile. The unoptimised SERS enhancement was approximately 2.5 × 10(4), thereby improving data acquisition time, reducing laser power requirements and enabling a citrate detection limit of 0.1 mM, which was well below the concentrations used in biofilm nutrient solutions. The flow confinement was observed by both optical microscopy and SERS imaging with good complementarity. We demonstrate the new bioreactor by growing flow-templated biofilms on the microchannel wall. This work opens the way for in situ spectral imaging of biofilms and their biochemical environment under dynamic flow conditions.

Aerobic biodegradation of 8:2 fluorotelomer stearate (FTS) and 8:2 fluorotelomer citrate triester (TBC) was evaluated in a forest soil in closed bottle microcosms. Loss of parent, production of 8:2 fluorotelomer alcohol (8:2 FTOH), which is released along with stearic acid (SA) by microbial ester linkage, and subsequent metabolites from FTOH degradation were monitored for up to 7months. Soil microcosms were extracted with ethyl acetate followed by two heated 90/10 v/v acetonitrile/200mM NaOH extractions. Cleavage of the ester linkage in the 8:2 FTS occurred (t1/2∼28d), producing 8:2 FTOH and various levels of subsequent metabolites. Quantifying the generation of SA from ester cleavage in FTS was complicated by the natural production and degradation of SA in soil, which was probed in an additional FTS and SA study with the same soil that had been stored at 4°C for 12months. In the latter study, FTS degraded faster (t1/2∼5d) such that SA production well above soil background levels was clearly observed along with rapid subsequent SA degradation. Cold storage was hypothesized to enrich fungal enzymes, which are known to be effective at hydrolytic cleavage. 8:2 TBC biotransformation was slow, but evident with the production of PFOA well above levels expected from known FTOH residuals. Slower degradation of TBC compared to FTS is likely due to steric hindrances arising from the close proximity of three 8:2 FT chains on the citrate backbone limiting the enzyme access., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

This study evaluated the effective dose of Ga-67 for a patient undergoing Ga-67 citrate nuclear examination by applying thermoluminescent dosimeter (TLD) technique and an indigenous water phantom. The Ga-67 radionuclide remaining in the body inevitably generated a measurable internal dose even though gamma camera scanning took only minutes to complete the clinical examination. For effective simulation of the cumulated effective dose for a patient undergoing examination, 150 TLDs were placed inside the water phantom for 6 days to monitor the gamma ray dose from the distributed Ga-67 citrate solution. The inserted TLDs represented internal organs, and the effective dose was calculated according to data in the ICRP-60 report. The water phantom was designed to model the body of a healthy human weighing 70 kg, and the water that was mixed with Ga-67 citrate solution was slowly replaced with fresh feed water to yield the required biological half life of the phantom. After continuously feeding in fresh water throughout the 6 days of TLD exposure, the TLDs were analyzed to determine the effective doses from the various biological half lives of the phantom. The derived effective dose of 185 MBq Ga-67 citrate solution for male/female (M/F) was 10.7/12.2, 10.7/12.0, 8.7/9.9 and 6.0/6.8 mSv, of biological half lives of 6.0, 4.5, 3.0 and 1.5 days, respectively. Although these experimental results correlated well with earlier empirical studies, they were lower than most calculated values. The cumulated uncertainty in the effective dose was 12.5-19.4%, which was acceptable in terms of both TLD counting statistic and reproducibility.

International normalized ratio (INR) discrepancies were noted between clinical laboratories using various prothrombin time (PT) systems. We studied the influence of different commercial blood collection tubes and different PT systems on INR measurements. INRs of fresh patient samples were determined by 3 laboratories, each using different PT systems. In the first part of the study, samples were drawn with Vacutainer tubes and in the second part with Monovette tubes. In the first part of the study, the maximum bias for all patients amounted to 0.46 INR (14%), and in the second part, to 0.14 INR (4.9%). The maximum bias for all patients could be reduced further by local system calibration using frozen pooled plasma specimens. The sodium citrate solutions in the blood collection tubes were contaminated with magnesium ions (approximately 2.7 mmol/L and 0.3 mmol/L in the Vacutainer and Monovette, respectively). INR discrepancies could be explained largely by this influence of blood collection tubes. The maximum allowable magnesium contamination in sodium citrate anticoagulant solutions should be less than 1 mmol/L.

Objectives: The aim of this study was to establish the parameters of concentration, time and mode of application of citric acid and sodium citrate in relation to root conditioning., Material and Methods: A total of 495 samples were obtained and equally distributed among 11 groups (5 for testing different concentrations of citric acid, 5 for testing different concentrations of sodium citrate and 1 control group). After laboratorial processing, the samples were analyzed under scanning electron microscopy. A previously calibrated and blind examiner evaluated micrographs of the samples. Non-parametric statistical analysis was performed to analyze the data obtained., Results: Brushing 25% citric acid for 3 min, promoted greater exposure of collagen fibers in comparison with the brushing of 1% citric acid for 1 minute and its topical application at 1% for 3 min. Sodium citrate exposed collagen fibers in a few number of samples., Conclusion: Despite the lack of statistical significance, better results for collagen exposure were obtained with brushing application of 25% citric acid for 3 min than with other application parameter. Sodium citrate produced a few number of samples with collagen exposure, so it is not indicated for root conditioning.

Purpose: Magnetic resonance stereoscopic imaging (MRSI) of the prostate is an emerging technique that may enhance targeting and assessment in radiotherapy. Current practices in radiotherapy invariably involve image guidance. Gold seed fiducial markers are often used to perform daily prostate localization. If MRSI is to be used in targeting prostate cancer and therapy assessment, the impact of gold seeds on MRSI must be investigated. The purpose of this study was to quantify the effects of gold seeds on the quality of MRSI data acquired in phantom experiments., Methods and Materials: A cylindrical plastic phantom with a spherical cavity 10 centimeters in diameter wss filled with water solution containing choline, creatine, and citrate. A gold seed fiducial marker was put near the center of the phantom mounted on a plastic stem. Spectra were acquired at 1.5 Tesla by use of a clinical MRSI sequence. The ratios of choline + creatine to citrate (CC/Ci) were compared in the presence and absence of gold seeds. Spectra in the vicinity of the gold seed were analyzed., Results: The maximum coefficient of variation of CC/Ci induced by the gold seed was found to be 10% in phantom experiments at 1.5 T., Conclusion: MRSI can be used in prostate radiotherapy in the presence of gold seed markers. Gold seeds cause small effects (in the order of the standard deviation) on the ratio of the metabolite's CC/Ci in the phantom study done on a 1.5-T scanner. It is expected that gold seed markers will have similar negligible effect on spectra from prostate patients. The maximum of 10% of variation in CC/Ci found in the phantom study also sets a limit on the threshold accuracy of CC/Ci values for deciding whether the tissue characterized by a local spectrum is considered malignant and whether it is a candidate for local boost in radiotherapy dose., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Plasticizers in 101 samples of polyvinyl chloride (PVC) toys on the Japanese market were surveyed. No phthalates were detected in designated toys, though bis(2-ethylhexyl)phthalate, diisononyl phthalate, diisobutyl phthalate, dibutyl phthalate, diisodecyl phthalate and benzyl butyl phthalate were detected in more than half of other toys. 2,2,4-Tributyl-1,3-pentanediol diisobutylate, o-acetyl tributyl citrate, adipates and diacetyl lauroyl glycerol, which are alternative plasticizers to phthalates, were detected. The results of structural analysis confirmed the presence of di(2-ethylhexyl)terephthalate, tributyl citrate, diisononyl 1,2-cyclohexanedicarboxylate and neopentyl glycol esters; these have not previonsly been reported in Japan. There appears to be a shift in plasticizers used for designated toys from phthalates to new plasticizers, and the number of different plasticizers is increasing.

Hydrophilic interaction chromatography (HILIC) has emerged as a very useful separation method for polar analytes, including non-covalent metal species. Several types of stationary phases are available for HILIC applications, differing mainly in their chemical functionalities that supply additional interaction modes and alternative selectivities for the separation of special analytes. With regard to the separation of metal species only few of these stationary phases have been applied to date, and it is not completely clear what are their differences with respect to the chromatographic separation of metal species, but also with respect to species stability during chromatography. Here, a comparison of different column types for the HILIC separation of iron citrate and copper histidine species is presented and the results are discussed with respect to retention mechanisms and chromatographic stability of these metal species. It is shown that different stationary phases display very different separation patterns. In particular, three types of HILIC columns enable successful separation of iron citrates and copper histidine at pH 5.5, namely a crosslinked diol phase, a zwitterionic phase, and an amide phase. Two groups of iron-citrates are separated on all three columns, consisting of a species of 3:3 stoichiometry and another one of mainly 3:4 stoichiometry (plus 1:2 and 2:2 species). For copper-histidine only one stable species is found based on the 1:2 stoichiometry. Detection and unambiguous identification of the different species is possible by employing electrospray mass spectrometry in the negative ionization mode. Species found in standard solutions are consistent with species found in spiked plant samples. Also in unspiked solutions iron citrate of 3:4 stoichiometry (plus 1:2 and 2:2) is detectable, but no species of 3:3 stoichiometry. Significant differences of related species patterns are found in real plant samples., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Migration levels of acetyl tributyl citrate (ATBC) plasticiser from polyvinyl chloride (PVC) film into the European Union specified aqueous food simulants (distilled water, 3% w/v acetic acid and 10% v/v ethanol) were monitored as a function of time. Migration testing was carried out at 40°C for 10 days. Determination of the analyte was performed by applying an analytical methodology based on surfactant (Triton X-114) mediated extraction prior to gas chromatographic-flame ionisation detection. PVC cling film used was subjected to ionising treatment with a [(60)Co] source, as well as to electron-beam irradiation at doses equal to 5, 15 and 25 kGy, with the aim to compare the effect of type and dose of radiation on the specific migration behaviour of PVC. Equilibrium concentrations of acetyl tributyl citrate into the aqueous solvents covered the ranges 173-422 µg l(-1) and 296-513 µg l(-1) for gamma- and electron-irradiated PVC, respectively. Hence, e-beam irradiation resulted in significantly higher ATBC migration compared with gamma treatment. The highest extraction efficiency of the 10% ethanol solution was common in both gamma and e-beam treatments; distilled water demonstrated the lowest migration. Gamma-irradiation at intermediate doses up to 5 kGy produced no statistically significant (p > 0.05) effect on ATBC migration into all three aqueous simulants; however, this does not apply for high-energy electrons. Both ionising treatments were similar in that they resulted in statistically significant (p < 0.05) differences in plasticiser migrating amounts between non-irradiated and irradiated at doses of 15 and 25 kGy samples. Gamma-radiation did not affect the kinetics of plasticiser migration. On the contrary, electron-beam radiation produced shorter equilibration times for all food-simulating solvents tested at 40°C. The above values regarding ATBC migration into aqueous food simulants are far below the European Union restriction (1 mg kg(-1) body weight) for both types of ionising radiation. Thus, PVC cling film may be used in food irradiation applications in contact with aqueous foodstuffs.

Unlabelled: Hibiscus cold (25 °C) and hot (90 °C) water extracts were prepared in various time-temperature combinations to determine equivalent extraction conditions regarding their physicochemical and phytochemical properties. Equivalent anthocyanins concentration was obtained at 25 °C for 240 min and 90 °C for 16 min. Total phenolics were better extracted with hot water that also resulted in a higher antioxidant capacity in these extracts. Similar polyphenolic profiles were observed between fresh and dried hibiscus extracts. Hibiscus acid and 2 derivatives were found in all extracts. Hydroxybenzoic acids, caffeoylquinic acids, flavonols, and anthocyanins constituted the polyphenolic compounds identified in hibiscus extracts. Two major anthocyanins were found in both cold and hot extracts: delphynidin-3-sambubioside and cyanidin-3-sambubioside. In general, both cold and hot extractions yielded similar phytochemical properties; however, under cold extraction, color degradation was significantly lower and extraction times were 15-fold longer., Practical Application: Hibiscus beverages are prepared from fresh or dried calyces by a hot extraction and pasteurized, which can change organoleptic, nutritional, and color attributes. Nonthermal technologies such as dense phase carbon dioxide may maintain their fresh-like color, flavor, and nutrients. This research compares the physicochemical and phytochemical changes resulting from a cold and hot extraction of fresh and dried hibiscus calyces and adds to the knowledge of work done on color, quality attributes, and antioxidant capacity of unique tropical products. In addition, the research shows how these changes could lead to alternative nonthermal processes for hibiscus.

Purpose: Glass transition temperature (T(g)) measurements of polymers are conventionally conducted in the dry state with little attention to the environment they are designed to work in. Our aim was to develop the novel use of dynamic mechanical analysis (DMA) to measure the T(g) of enteric polymethacrylic acid methylmethacrylate (Eudragit L and S) polymer films formulated with a range of plasticizers in the dry and wet (while immersed in simulated gastric media) states., Methods: Polymer films were fabricated with and without different plasticizers (triacetin, acetyl triethyl citrate, triethyl citrate, polyethylene glycol, propylene glycol, dibutyl phthalate, dibutyl sebacate). T(g) was measured by a dynamic oscillating force with simultaneous heating at 1 °C/min. This was conducted on films in the dry state and while immersed in 0.1M HCl to simulate the pH environment in the stomach., Results: The T(g) of unplasticized Eudragit L and S films in the dry state was measured to be 150 and 120 °C, respectively. These values were drastically reduced in the wet state to 20 and 71 °C for Eudragit L and S films, respectively. The plasticized films showed similar falls in T(g) in the wet state. The fall in T(g) of Eudragit L films to below body temperature will have far-reaching implications on polymer functionality and drug release., Conclusions: Immersion DMA provides a robust method for measuring T(g) of polymer films in the wet state. This allows better prediction of polymer behaviour in vivo., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Lactoferrin (Lf) is a molecule naturally present in bovine milk that affects the availability and transport systems of iron. Lf also binds endotoxin (lipopolysaccharide, LPS) of Gram-negative bacteria and modulates the immunological response. In the present study, concentrations of bovine Lf (bLf) and citrate in milk were determined in early (EL) and late (LL) lactating dairy cows, using an experimentally induced endotoxin mastitis model and a crossover design. Nine clinically healthy Finnish Ayrshire cows were challenged twice with 100 μg endotoxin infused into one udder quarter. Milk samples were collected from the challenged and control quarters of each cow before and after endotoxin infusion during 3 d, and bLf and citrate concentrations were measured. In all cows, clinical signs of mastitis were seen at both times of challenge, but the response was more severe in EL than in LL. Concentration of bLf in the milk started to rise approximately 8 h after endotoxin infusion and was still higher than normal on the third day, especially in the late-lactating cows. In milk of the LL group, concentrations of bLf were significantly higher than in the EL group. In contrast, concentrations of citrate were higher in milk of the EL cows compared with the LL cows. Concentration of bLf and citrate varied substantially among cows. The molar ratio of citrate to bLf before and after challenge was significantly higher during the EL period. The results of this study partly explain why cows in early lactation are more susceptible to intramammary infections and why mastitis is more severe in them.

Purpose: Citrate is a known inhibitor of calcium stone formation. Dietary citrate and alkali intake may have an effect on citraturia. Increasing alkali intake also increases urine pH, which can help prevent uric acid stones. We determined citrate, malate and total alkali concentrations in commonly consumed diet sodas to help direct dietary recommendations in patients with hypocitraturic calcium or uric acid nephrolithiasis., Materials and Methods: Citrate and malate were measured in a lemonade beverage commonly used to treat hypocitraturic calcium nephrolithiasis and in 15 diet sodas. Anions were measured by ion chromatography. The pH of each beverage was measured to allow calculation of the unprotonated anion concentration using the known pK of citric and malic acid. Total alkali equivalents were calculated for each beverage. Statistical analysis was done using Pearson's correlation coefficient., Results: Several sodas contained an amount of citrate equal to or greater than that of alkali and total alkali as a lemonade beverage commonly used to treat hypocitraturic calcium nephrolithiasis (6.30 mEq/l citrate as alkali and 6.30 as total alkali). These sodas were Diet Sunkist Orange, Diet 7Up, Sprite Zero, Diet Canada Dry Ginger Ale, Sierra Mist Free, Diet Orange Crush, Fresca and Diet Mountain Dew. Colas, including Caffeine Free Diet Coke, Coke Zero, Caffeine Free Diet Pepsi and Diet Coke with Lime, had the lowest total alkali (less than 1.0 mEq/l). There was no significant correlation between beverage pH and total alkali content., Conclusions: Several commonly consumed diet sodas contain moderate amounts of citrate as alkali and total alkali. This information is helpful for dietary recommendations in patients with calcium nephrolithiasis, specifically those with hypocitraturia. It may also be useful in patients with low urine pH and uric acid stones. Beverage malate content is also important since malate ingestion increases the total alkali delivered, which in turn augments citraturia and increases urine pH., (Copyright 2010 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.)

The phenolic fraction and other polar compounds of the Hibiscus sabdariffa were separated and identified by HPLC with diode array detection coupled to electrospray TOF and IT tandem MS (DAD-HPLC-ESI-TOF-MS and IT-MS). The H. sabdariffa aqueous extract was filtered and directly injected into the LC system. The analysis of the compounds was carried out by RP HPLC coupled to DAD and TOF-MS in order to obtain molecular formula and exact mass. Posterior analyses with IT-MS were performed and the fragmentation pattern and confirmation of the structures were achieved. The H. sabdariffa samples were successfully analyzed in positive and negative ionization modes with two optimized linear gradients. In positive mode, the two most representative anthocyanins and other compounds were identified whereas the phenolic fraction, hydroxycitric acid and its lactone were identified using the negative ionization mode.

Sodium caprylate and N-acetyltryptophan are the most frequently used stabilizers that protect the albumin from aggregation or heat induced denaturation. In turn citrates - excipients remaining after fractionation process - can be treated as by-product favoring leaching aluminum out of glass containers whilst albumin solution is stored. With ionic nature these substances have all the markings of a subject for capillary electrophoresis analysis. Thus CE methods were proposed as new approach for quality control of human albumin solution in terms of determination of stabilizers and citrates residue. Human albumin solutions both 5% and 20% from various manufacturers were tested. Indirect detection mode was set to provide sufficient detectability of analytes lacking of chromophores. As being anions analytes were separated with reversed electroosmotic flow. As a result of method optimization two background electrolytes based on p-hydroxybenzoic acid and 2,6-pyridinedicarboxylic acid were selected for stabilizers and citrates separation, respectively. The optimized methods were successfully validated. For citrates that require quantification below 100microM the method demonstrated the precision less than 4% and the limit of detection at 4microM. In order to check the new methods accuracy and applicability the samples were additionally tested with selected reference methods. The proposed methods allow reliable quantification of stabilizers and citrates in human albumin solution that was confirmed by method validation as well as result comparison with reference methods. The CE methods are considered to be suitable for quality control yet simplifying and reducing cost of analysis.

Citrate complexes of Mn and Fe, and potentially those of Cu and Zn, are considered as important low molecular mass species in human serum and cerebrospinal fluid (CSF). For example, Mn is supposed to enter the brain under excess exposure as Mn-citrate leading to neurotoxic effects. Mn-citrate has been characterised in human CSF using chromatography and electrophoresis online with inductively coupled plasma mass spectrometry, but not yet with molecular mass spectrometry. Therefore, this study explores the potential of electrospray ionisation (ESI) with selected reaction monitoring (SRM) for the detection of metal-citrate complexes, in particular Mn-citrate. The collision-induced dissociation of precursor ions with various metal:citrate stoichiometries was studied for Mn-citrate, Fe-citrate, Cu-citrate and Zn-citrate. High selectivity was achieved for Mn(II)-citrate even in respect to Fe(III)-citrate which forms isobaric precursor ions. The limit of detection for Mn-citrate was estimated to be around 250 microg L(-1) (referring to the total Mn content in the standard) using flow injection. The sensitivity was sufficient for the determination of Mn-citrate in standard solutions and in an extract of an Mn-citrate-containing supplement. An improved ESI source design is expected to reduce the limits of detection significantly. The developed ESI-SRM method has the potential to provide complementary data for the quality control of current separation methods for metal citrates using element-selective detection, with application to biomedical samples and further matrices., (Copyright (c) 2009 John Wiley & Sons, Ltd.)